A NEW POLARIMETRIC METHOD FOR THE ANALYSIS OF DEXTRAN AND SUCROSE
J. D. Miller USDA-ARS Sugarcane Field Station
Canal Point, Florida 33438
Variety maturity tests were conducted on 16 Canal Point (CP) clones at 5 locations over 3 years in the Everglades Agricultural Area in Florida. Cane sugar quality was measured at biweekly intervals during the October to March harvest season in each year. A quadratic response function of lbs. sucrose per gross ton of cane (SPT) vs. sampling date was calculated for each clone using the entire 3-year data set, and date and magnitude of maximum SPT calculated. CP89-2143 and CP72-2086 had the highest predicted SPT at 305 and 285 on Feb 9 and Feb 13, respectively. Model fit varied greatly between clones, with R2 values ranging from 0.23 - 0.72. In general, clones with higher R2 values tended to have maximum SPT after February 1. The SPT data was then divided into "early", "middle", and "late" maturity classes and the CP clones ranked based on average SPT within a given class. Results of this analysis will be discussed in terms of a harvest scheduling aid for Florida growers.
Journal American Society of Sugarcane Technologists, Vol. 2 2 , 2 0 0 2
Protox Inhibitor Herbicide Effects on Pythium and Root Rot of Sugarcane J. H. Daugrois
Cirad-ca, Sugarcane Program
Station de Roujol, Guadeloupe, 97170 Petit Bourg, FWI J. W. Hoy and J. L. Griffin
Department of Plant Pathology and Crop Physiology LSU AgCenter, Baton Rouge, LA 70803
A complex of root pathogens contributes to yield decline of sugarcane. Pythium root rot, caused by P. arrhenomanes, is one component of the disease complex. Root rot control would increase yield and could allow additional ratoons to be obtained. Herbicides can have non-target effects, such as enhancing or reducing root disease severity. Protoporphyrinogen oxidase (protox) inhibitor herbicides may reduce fungal disease severity in other crops by inducing host resistance. In addition, visual growth increases in sugarcane early growth following application of one protox inhibitor herbicide have been observed. Therefore, lab and greenhouse experiments were conducted to determine protox inhibitor herbicide effects on Pythium, root rot severity, and sugarcane growth.
Three protox inhibitor herbicides, Milestone (azafeniden), Spartan (sulfentrazone), and Valor (flumioxazin) were evaluated for their effects on in vitro mycelial growth rate of P.
arrhenomanes, P. ultimum, and P. aphanidermatum and Pythium root rot and growth of sugarcane in two greenhouse experiments. Effects on sugarcane growth and root rot were evaluated after herbicide leaf or soil application at the recommended rate and 1/10 and 1/20 the recommended rate. Three types of soil were used, field soil (FS), sterilized field soil (SFS), and sterilized field soil infested with P. arrhenomanes (SFS+P).
All three herbicides strongly reduced Pythium mycelial growth in vitro. No growth of P.
arrhenomanes occurred when rate one or above was applied in the growth medium. Mycelial growth inhibition still occurred at a 200-fold dilution of the recommended rate. Milestone had the strongest effect followed by Spartan and Valor. In the greenhouse, all three herbicides reduced P, arrhenomanes root colonization in some cases, but results were erratic between experiments. Milestone and Valor were phytotoxic in sterile and nonsterile soils, and with a short duration experiment, the damage may have made it difficult to detect effects on root rot severity and plant growth. No treatment clearly reduced visual root rot symptoms. Only 1/10 rate Spartan applied to leaves significantly reduced P. arrhenomanes colonization in SFS+P and increased plant growth. In field soil, more treatments reduced Pythium root colonization, but only leaf-applied Spartan at rate one and 1/10 rate Valor increased some component of sugarcane growth.
No consistent effects on disease severity and plant growth were shown. However, the greenhouse experimental system may not have been sufficient to clearly demonstrate the effects of the protox inhibitor herbicides on sugarcane root rot. Although variable, the results suggest these herbicides may be capable of reducing P. arrhenomanes infection and increasing plant growth through reduced root rot severity. The slight increases in plant growth following leaf application of herbicide suggest an indirect effect through induced resistance.
Journal American Society of Sugarcane Technologists, V o l 22,2002
Irrigation of Sugarcane on Clay in a High-Rainfall Environment Howard P. Viator
Iberia Research Station LSU AgCenter
Variable yield responses to irrigation of sugarcane, Saccharum spp., in Louisiana's humid climate have made it difficult to evaluate its economic soundness. Nevertheless, the occurrence of several droughts during the past decade in southern Louisiana has intensified the interest in supplemental irrigation. During the severe drought of 2000, a study to evaluate the response of LCP 85-384 plant cane to irrigation was conducted on an Alligator clay soil (thermic Vertic Haplaquept), a soil textural class that tends to restrict root development under drought conditions. Irrigation was scheduled when stalks elongated 5 cm or less per week.
Supplemental water was supplied in furrows on May 5, May 25, July 21 and August 28 for a cumulative total of 1130 m3 . The experimental site received a total of only 50.5 cm of rain from May through October, a rainfall deficit of 38.4 cm when compared to a 25-yr average for the same period. Height difference at harvest between the irrigated and non-irrigated plots was 50 cm. Yields mirrored the plant height disparity, with irrigated plots producing 44% higher cane (P = .06) and sugar (P = .08) yields than the control plots. The magnitude of the yield responses to irrigation in this experiment, 22.6 Mg ha-1 of cane and 2.41 Mg ha-1 of sugar, was comparable to that observed elsewhere under similar dry conditions.
Effect of Tissue Culture Method on Sugarcane Yield Compnents J. W. Hoy
Department of Plant Pathology and Crop Physiology LSU AgCenter, Baton Rouge, LA 70803
K. P. Bischoff and K. A. Gravois Sugar Research Station LSU AgCenter, St. Gabriel, LA 70776
S. B. Milligan United States Sugar Corporation
Clewiston, FL 33440
Vegetative propagation is conducive to the spread of systemic sugarcane diseases, such as ratoon stunting disease (RSD). This important disease is now controlled in Louisiana largely by planting commercial seed-cane initially produced through tissue culture. Kleentek®
seed-cane has been available to farmers since the late 1980s. In the early years, farmers sometimes noted that tissue culture derived plants had smaller stalk diameter and weight and a higher stalk population. The tissue culture method used at that time was leaf roll callus culture.
Since then, the method has been changed to direct regeneration from the apical meristem to
Journal American Society of Sugarcane Technologists, Vol. 2 2 , 2 0 0 2
attempt to reduce or eliminate differences between tissue culture derived plants and the original varieties.
To determine whether tissue culture method affects yield or its components, three varieties, CP 70-321, LCP 85-384, and HoCP 85-845, were compared in three successive crops, plant cane through second ratoon, at three locations. Experiments were planted with stalks from three sources: Kleentek plants derived from callus (undifferentiated cells) produced from the leal roll above the apical meristem, Kleentek plants directly regenerated from an apical meristem, and original plants from conventional bud propagation. Stalks of plants derived from both tissue culture methods were typical of Kleentek seed-cane farmers would purchase for planting that had been rogued for phenotypic variants (off-types) and increased by bud propagation. Yield components compared included stalk diameter, length, weight, sucrose content, and population;
cane tonnage; and sugar yield. Plants were visually inspected for off-types in May, August, and at harvest.
Differences in yield components between the two tissue culture methods and bud- propagated cane only occurred in CP 70-321. Stalk diameter and stalk weight were lower and stalk population was higher for plants derived from leaf roll callus compared to bud propagated cane. However, all yield components were similar for plants derived from apical meristem and bud propagation. Individual plant off-types were not observed in cane produced by either tissue culture method. In summary, variety and tissue culture method affected persistent, uniform variation in plant growth habit resulting from tissue culture that changed some yield components. However, apical meristem culture was suitable for production of seed-cane, as sugarcane derived by meristem culture of all three varieties did not differ significantly from the original germplasm for any measured trait.
Genes Expressed During Regeneration in Tissue Culture Robin Rowe
University of New Orleans Candace Timple and Sarah Lingle
USDA-ARS-SRRC
Regeneration from tissue culture by way of somatic embryogencis is common in many varieties of sugarcane, but many economically important varieties of sugarcane are recalcitrant.
Better understanding of the genetic control of embryogenesis could lead to the ability to transfer this trait to important varieties lacking it. This could assist in the rapid progation of these varieties and in the construction of beneficial transgenic varieties. We used differential display techniques to compare genes expressed in mRNA samples from non-cmbryogenic, proembryogenic, and embryogenic callus from variety CP 72-1210 and from non-embryogemc callus from the recalcitrant variety TCP 87-3388. Several novel sequences were identified. One codes for a hypothetical protein containing several phosphorylation sites. Another codes for a hypothetical protein with a glycosylation site and a camp controlled phosphorylation site. The
Journal American Society of Sugarcane Technologists, Vol. 22,2002
third codes for a hypothetical protein with a 37% homology to extension in canola. The last codes for a hypothetical protein that has a 93% homology to a putative glucose-6- phosphate/phosphate translocator in rice. Whether these sequences are unique to a specific tissue type is still under investigation.
A Technique to Breed for Ratoon Stunting Disease in Sugarcane J. D, Miller, J. C. Comstock, P,Y. P. Tai, and B. Glaz
USDA-ARS Sugarcane Research Station Canal Point, Florida
Ratoon stunting disease (RSD) caused by Clavibacter xyli subsp. xyli is one of the most important sugarcane (interspecific hybrids of Saccharum spp.) diseases in Florida. The objective of this study was to evaluate the effectiveness of stubble inoculation and determine if it could be used in a program to breed for RSD resistance. Field grown seedling sugarcane plants were inoculated at maturity by cutting with knives dipped in juice infected with ratoon stunting disease bacteria (RSD). The regrowth from these stools was sampled at the base of the mature stalks and RSD susceptibility was based on the number of colonized vascular bundles determined using the tissue blot immunoassay. After selection based on vegetative characteristics in Seedlings, the average RSD rating of 12 crosses with 658 selections was 1.52.
When resampled as mature plants in Stage I, the average rating was 4.15. The plants were reinoculated and replanted into a Stage 1 sized plot. There were 67 clones selected for advancement to Stage II. They had an average RSD rating of 1.75. One major advantage of this system is that it requires no special planting in which to evaluate RSD resistance. The major disadvantage of this system from our standpoint in Florida is that it requires that seedling selection be done in the ratoon crop and that all clones in the breeding program would potentially be infected with RSD. In all probability very high yielding susceptible clones would be dropped with this selection scheme. Growers in Florida now manage RSD with a combination of genetic resistance and clean seed cane. Therefore, our industry is not willing to lose those potentially high yielding clones that are susceptible but could be profitable when grown without RSD.
Progress in the Development of Transgenic Disease-Resistant Sugarcane Z. Ying and M. J. Davis
University of Florida, Tropical Research and Education Center Homestead, Florida
Efforts are underway to develop sugarcane with transgenic resistance to the sugarcane yellow leaf luteovirus (SCYLV), leaf scald disease (LSD), and ratoon stunting disease (RSD).
Genetic constructs containing the SCYLV coat protein in the sense (pFM395) and antisense (pFM396) orientations were obtain from T. E. Mirkov (Texas A&M, Weslaco). A genetic construct (pMBP39-22) containing a modified cecropin gene (MB39) was obtained from Lowell
Journal American Society of Sugarcane Technologists, Vol. 22,20O2
Owens (USDA, Beltsville, MD), to vitro growth inhibition assays indicated that MB39 should be highly active against the RSD and LSD pathogens, Clavibacter xyli subsp. xyli and Xanthomonas albilineans, respectively. A number of other DNA constructs were made including those with the cecropin gene under control of the maize ubiquitan promoter (pZY-C), and the antisense SCYLV gene fused with the cecropin gene both under control of the ubiquitan promoter (pZY-CSA). Sugarcane callus cultures were co-bombarded with the individual constructs and another construct containing the NPT II gene as a selectable marker. Genetically transformed plants were regenerated from these materials and are being tested further.
Potential Impact of DNA Marker Technology on Sugarcane Breeding Yong-Bao Pan
USDA-ARS, Southern Regional Research Center, Sugarcane Research Unit, 5883 USDA Road, Houma, LA 70360, U.S.A.
At the turn of the new millennium, breeders have begun to realize how DNA marker technology may potentially impact traditional sugarcane breeding programs. Sugarcane is a tropical grass with both male and female organs within each tiny flower. Self-pollination may occur even after a male-sterility treatment such as the immersion of tassels in hot water or alcohol. The use of DNA marker technology may allow breeders to eliminate progeny from unwanted selfs early in the basic and commercial programs. At least five classes of DNA markers are available to use, each having its strong and weak points. These are restriction fragment length polymorphism (RFLP), random amplified polymorphic DNA (RAPD), polymerase chain reaction (PCR), simple sequence repeat (SSR) or microsatellites, and amplified fragment length polymorphism (AFLP). Unlike the morphological traits, DNA fingerprints constructed with these classes of markers are quite reliable and not influenced by the environment. A few PCR {Eri3IEH4 and GigllPII), RAPD (OPA11-366), and SSR (SMC334BS, SMC336BS and MCSA068G08) markers, that prove to be species-specific, have been developed to assist in the basic selection program at the Sugarcane Research Unit at Houma, Louisiana.
Multi-disciplinary studies are underway to identify and clone RAPD or AFLP markers that are tightly linked to genes contributing to important agronomic traits. Multi-institutional collaborations are also being sought to construct microsatellite linkage maps from several genetic populations (Fl, F2, BC1) of sugarcane.
In Vivo Viability Assay of Sugarcane Pollen Stored at Ultra Low Temperature Following Preservation Treatments
P. Y. P. TaiandJ. D.Miller USDA-ARS Sugarcane Field Station
Canal Point, Florida
Storage of sugarcane pollen is desirable for enhancing germplasm because of the different flowering time. The viability of Saccharum spontaneum pollen can be significantly prolonged under low temperature after being properly air dried to reduce its moisture content.
Journal American Society of Sugarcane Technologists, Vol. 22,2002
The information on pollen viability of commercial cultivars (CP 70-1133, CP 98-1301, and CP 98-1654) were used to examine their viability after being stored at low temperature. Pollen samples were collected in the early morning after anthesis and divided into two sets: the first was dried in a cool dehumidified room for three hours and the second set was treated with cryoprotectants. Both sets of pollen were stored immediately at -8G°C for 1 to 4 months.
Cryoprotectants included 0.25 - 0.5 M solutions in various combinations of dimenthyl sulfoxide, glycerol, sorbitol, and sucrose. An in vivo assay was used to measure the pollen viability.
Pollen was applied onto the tassels of green canes, CP 65-357 and Green German (S.
officinarum), in the morning during the flowering season. Fuzz was harvested about 30 days after pollination for germination test. Seedlings were transplanted to field. Seedlings from crosses derived from stored S. officinarum pollen were classified based on the gross plant morphology at 4-month-old while seedlings derived from crosses with stored pollen of commercial cultivars were classified based on stalk colors. Stalk color was determined by one intemode from each of 12-month-old seedlings that was cut and dipped vertically in 5%
sulfurous acid solution for 3-4 days to eliminate chlorophyll pigment. Loss of pollen viability (%) due to preservation treatments was estimated by [1 - (seed set from stored pollen)/(seed set from fresh pollen)] 100. Results showed that pollen of neither S. spontaneum nor commercial cultivars produced viable seedlings when they were stored at -80°C after being treated with cryoprotectants. After being exposed to air drying, pollen of both S. spontaneum and commercial cultivars produced viable seedlings ranging from poor to good seed set when the stored pollen was used to cross with CP 65-357 or Green German. Average losses of pollen viability were 50% (1997/98) and 88% (1999/00) for CP 98-1654. In addition to the use of the pollen storage for germplasm enhancement, this study suggests that stored pollen with genetic marker may be used to help identify hybrids for genetic and breeding investigations.
Journal American Society of Sugarcane Technologists, V o l 2 2 , 2 0 0 2
MANUFACTURING ABSTRACTS
The Freeze of 2001-A "New Book is Written"
John A. Fanjul Atlantic Sugar Associations, Inc.
Belle Glade, Florida
Atlantic Sugar Associations, Inc. developed an organizational plan, which involved pooling its R&D/Harvesting, Operations/Mill, and Cane Bank, to handle the freeze in 2001.
Atlantic Sugar Associations, Inc. had successful and record-breaking results across the board.
The Breakage in Sugarcane Mill Rolls Jorge Okhuysen
Mexico
The causes of failure involving the design, materials selection, methods of manufacturing, and the influence of operating conditions in sugarcane mill rolls will be discussed.
Material Balance and Equipment Requirements of a Typical Sugar Mill Eduardo Samour, P.E. and William Easdaie
United States Sugar Corporation Clewiston, FL
Traditionally, to reduce production costs or for other reasons, most sugar mills have increased their grinding rate over the years, after they were designed and built for certain capacity, and conditions. When an expansion project is conceived in a sugar mill, the focus generally is, on cane grinding capacity and steam production. Even though these are extremely important factors, a proper evaluation of the rest of the equipment in the factory is often neglected. This, bring about unnecessary bottlenecks that will defeat the purpose of the expansion, or even worse, a reduction of efficiency. With a properly conducted survey of equipment capacities, an engineer can determine, with the new operating conditions, the proper capacity required in each station of the process.
This paper describes, calculations of material and steam balance performed for a typical sugar mill. It is based on a grinding rate of 1000 tons of cane per day, using the double magma system, and quadruple effect evaporation, with first effect vapor bleeding for secondary heaters and clarified juice heaters and second effect vapor bleeding for primary heaters and vacuum pans.
Journal American Society of Sugarcane Technologists, Vol. 22,2002
The results are presented in various charts. These were developed, to illustrate different volumes of materials that can be expected in the boiling house, under different cane quality conditions. Other charts are also presented such as: heating surface required for Juice heaters on the various stages, evaporation rates necessary to satisfy the demands of vacuum pans, and heaters. These figures are useful for sizing the proper equipment required under different conditions and grinding rate.
Properly planning an expansion project, after evaluating all the areas of the mill, will help mill managers spend their investment dollars in the areas were equipment is most needed.
A properly balanced factory, provides a smooth operation that enable the mill engineers to focus their attention on increasing efficiency, rather than coping with the added material they have to process.
Reducing Equipment Cost/Best Equipment Management Practices Neal Hahn
Nortrax Equipment Company - South Baton Rouge, Louisiana
The owning and operating cost of mobile equipment can have an adverse effect on a mill's profitability. Cost control is important. The core business of the mill is grinding cane, rather than mobile equipment management. Many managers do not take the time to consider this key area of operation. The productivity of equipment is directly proportional to the effectiveness of an equipment management strategy. Equipment that stays idle during productive times is a substantial cost to the mill. Utilization tracking can be used to determine if added equipment is required. Downtime can be an indicator both of equipment and maintenance problems. A good program of maintenance for high-tech equipment must include oil sampling, repair option management, preventative maintenance, and life cycle planning. A good record keeping system should also include an effort to make historical comparisons of cost per hour. The equipment division of each mill should also have a Standard Operating Procedures guide, which would address the key areas of equipment operation and maintenance. This paper will provide ideas on better equipment management and review specific examples key to lowering the operating cost of equipment.
What You Should Learn from Your Chemical Supplier Stephen J. Clarke
Florida Crystals Corporation
This paper surveys the issues of selection, use and fate of chemicals used as processing aids in sugar production and in equipment cleaning. The chemical sales business is extremely competitive and it is essential that the sugar technologist (chemical user) be aware of the benefits, costs, and possible unforeseen consequences of each chemical used. The chemical supplier who should be familiar with the scientific basis for the application must provide this information - there is no magic in this business. Chemical use should be minimal but is